General tips
- mix all samples with 10-20µl of PBS, else the sperm may clump and stick to each other making measurements of single cells harder
- breeding status of the bird does not matter (some say that very early and very late breeding season should be avoided)
- if sacrificing birds, take testes and vas deference and use those to prepare fresh microscope slides (see below) and to store for later
- slides with sperm can be air-dried, in general no stains needed
Equipment
- phosphate buffered saline solution (PBS): mix one PBS tablet (Sigma; product number P-4417) with 200ml water (use gloves, if you touch the tablet)
- eppies with 100µl of 5% formalin
- pipette tips (use a new pipet tip for each bird/wash)
- microscope slides
- dissecting tools (forceps, scalpel, dissecting microscope)
- if staining, then (KIM)
- microscope and camera set up for sample inspection and photographing
Sperm collection
Abdominal massage
- pipette 10 µl of PBS into the lid of an eppi with formalin,
- massage the cloaca